Purification and characterization of 3':5'-cyclic GMP-dependent protein kinase.

摘要: Abstract Guanosine 3':5'-cyclic monophosphate (cGMP)-dependent protein kinase has been purified to homogeneity from bovine lung by affinity chromatography and characterized. Partially kinase, specifically activated low concentrations of cGMP (22 NM), was adsorbed onto 8-(2-aminoethyl)-amino-adenosine monophosphate-Sepharose. After washing remove nonspecific proteins, cGMP-dependent eluted 0.1 mM cGMP. The contained specific binding activities. Purification the holoenzyme possible because subunit dissociation does not occur upon cyclic nucleotide binding. an apparent molecular weight 150,000 as determined glycerol density gradient sedimentation. On sodium dodecyl sulfate/polyacrylamide gel electrophoresis, a single band 71,000 observed that suggested is dimer composed subunits identical weight. required high Mg+2 for optimal activity; heat-stable modulator which inhibited adenosine monophosphate-dependent activity had no effect on kinase.